Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
  • A01G7/00—Botany in general
  • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
  • A01N31/06—Oxygen or sulfur directly attached to a cycloaliphatic ring system
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
  • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
  • A01N63/38—Trichoderma
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
  • A01P21/00—Plant growth regulators

Definitions

• the present invention relates to an agent for improving environmental stress resistance.
• plants can encounter various environmental stresses, such as high temperatures, low temperatures, dryness, excessive humidity, low light, and salt. Unlike animals, plants cannot move quickly to escape stress, so they have developed mechanisms to adapt to these environmental stresses.
• Such methods include (1) crossbreeding, (2) recombination of environmental stress resistance genes (e.g., Patent Document 1), and (3) chemical treatment methods (e.g., Patent Documents 2 and 3).
• Patent Document 1 describes the introduction of glutathione S-transferase (GST) gene into rice to impart low temperature stress tolerance.
• GST glutathione S-transferase
• gene recombination has a problem of low social acceptance.
• the method of treating the above-mentioned (3) compound depending on the type of compound, there are concerns about safety to the human body and the environment.
• sanguinarine is used in Patent Document 2
• zerumbone is used in Patent Document 3
• the sanguinarine described in Patent Document 2 is a toxic alkaloid.
• Non-Patent Document 1 a growth-repression phenotype
• Patent Document 4 describes a composition for improving abiotic stress resistance, which contains a red pepper extract extracted from the flesh of red pepper as an active ingredient and has an effect of improving stress resistance to one or more types selected from the group consisting of temperature stress, chemical stress, light stress, dry stress, pH stress, salt stress, hypoxic stress, herbicide stress, physical stress, and disease stress.
• Example 1 of the same document specifically states that "1 kg of red pepper was decapitated, cut into 1 cm cubes, and crushed in a mixer. Impurities were removed by filtration using filter cloth and filter paper. The obtained filtrate was concentrated about 10 times using a rotary evaporator to obtain a concentrate.
• the present invention aims to provide an environmental stress resistance enhancer that can improve the environmental stress resistance of plants without interfering with normal plant growth.
• the present invention is as follows.
• Item 1 An agent for improving environmental stress resistance for plants, comprising capsanthin and/or its fatty acid ester.
• Item 2. Item 2. The environmental stress tolerance improving agent according to Item 1, wherein the capsanthin and/or fatty acid ester thereof is derived from a plant of the genus Capsicum in the family Solanaceae.
• Item 3. The capsanthin and/or its fatty acid ester is an extract extracted from at least one plant selected from the group consisting of red paprika, red chili pepper, and red bell pepper; and Item 2. The agent for improving environmental stress resistance according to Item 1, wherein the extract is hydrophobic.
• Item 4. Item 2. The agent for improving environmental stress resistance according to Item 1, wherein the environmental stress resistance is high temperature stress resistance.
• Item 2. The agent for improving environmental stress resistance according to Item 1, wherein the environmental stress resistance is drought stress resistance.
• Section 6. Item 2.
• Item 7. Item 7.
• Section 8. Item 8.
• An environmental stress tolerance improver composition comprising the environmental stress tolerance improver according to any one of Items 1 to 7, and further comprising a fertilizer component and/or an additive.
• Item 9. Item 8.
• a method for improving environmental stress tolerance of a plant comprising applying the environmental stress tolerance improver according to any one of Items 1 to 7, or the environmental stress tolerance improver composition according to Item 8 to a plant, or to soil or culture solution in which the plant grows.
• Item 10. Use of capsanthin and/or its fatty acid ester for improving environmental stress tolerance in plants.
• Item 11. A method for improving environmental stress tolerance in a plant, comprising: 1. A method comprising applying capsanthin and/or its fatty acid esters to a plant.
• Item 12. The method according to Item 11, wherein the improved environmental stress resistance is an increase in the expression of a gene related to environmental stress resistance.
• Item 14. A method for producing a plant body, comprising the steps of: A method comprising applying capsanthin and/or a fatty acid ester thereof to a plant, cultivating the plant, and harvesting the plant.
• Item 15. A plant having improved environmental stress resistance obtained by applying capsanthin and/or a fatty acid ester thereof to the plant and cultivating the plant.
• Section 16 A gene expression promoter for promoting the expression of environmental stress resistance genes in plants, comprising capsanthin and/or a fatty acid ester thereof.
• Item 18. A composition for improving environmental stress tolerance in a plant, comprising capsanthin and/or a fatty acid ester thereof.
• Item 19. The composition according to item 18, wherein the plant is a Gramineae plant, a Solanaceae plant, a Cruciferae plant, a Leguminosae plant, an Amaranthaceae plant, or a Rosaceae plant.
• the composition according to item 18 or 19, wherein the environmental stress is at least one stress selected from the group consisting of high temperature stress, low temperature stress, oxidative stress, high light stress, drought stress, chemical stress, and injury stress.
• Item 22. A method for producing a plant having environmental stress tolerance, comprising applying capsanthin and/or a fatty acid ester thereof to the plant.
• Item 23. Item 23. The method according to Item 22, wherein the plant is a Gramineae plant, a Solanaceae plant, a Cruciferae plant, a Leguminosae plant, an Amaranthaceae plant, or a Rosaceae plant.
• Item 24 The method according to Item 22 or 23, wherein the environmental stress is at least one stress selected from the group consisting of high temperature stress, low temperature stress, oxidative stress, high light stress, drought stress, chemical stress, and injury stress.
• Section 25 The method according to Item 22 or 23, wherein the environmental stress is high temperature stress, drought stress, or salt stress.
• Section 26 A method for imparting environmental stress resistance to a plant, comprising applying capsanthin and/or a fatty acid ester thereof to the plant.
• the present invention provides an environmental stress resistance enhancer that can improve the environmental stress resistance of plants without interfering with normal plant growth.
• FIG. 1 shows a photograph (FIG. 1a) showing the condition of tomato seedlings after the tests described in Comparative Example 11 and Example 9 (paprika dye), and a photograph (FIG. 1b) showing the condition of tomato seedlings after the tests described in Comparative Example 11 and Reference Example 8 (indigo dye).
• the environmental stress tolerance improver of the present invention (the environmental stress tolerance improver can also be referred to as a "plant environmental stress tolerance agent” or a “plant stress tolerance imparting agent”) contains capsanthin and/or its fatty acid ester as an active ingredient.
• capsanthin and/or its fatty acid ester means only capsanthin, only a fatty acid ester of capsanthin, and a mixture of capsanthin and a fatty acid ester of capsanthin.
• the environmental stress resistance improver of the present invention may contain only capsanthin, may contain only a fatty acid ester of capsanthin, or may contain capsanthin and a fatty acid ester of capsanthin.
• the environmental stress resistance improver of the present invention may contain components other than capsanthin and/or its fatty acid ester.
• Capsanthin and/or its fatty acid esters are natural products contained in edible plants, and therefore are highly safe for the human body and the environment.
• capsanthin and/or its fatty acid esters are expensive, and therefore their use in agricultural applications is not practical. Therefore, the present inventors focused on paprika coloring, which is produced from paprika containing capsanthin and/or its fatty acid esters, is inexpensive, and is widely used as a food additive.
• Capsanthin is a natural red pigment that is a type of xanthophyll.
• Capsanthin or its fatty acid ester is a compound represented by the following general formula (I).
• R1 and R2 are the same or different and each represents a hydrogen atom or an acyl group derived from a fatty acid.
• capsanthin is a compound having a chemical structure in which R 1 and R 2 in the above general formula (I) are both hydrogen atoms (H).
• Capsanthin fatty acid esters include monoesters in which either one of R1 and R2 in the above general formula (I) is bonded with an acyl group derived from a fatty acid instead of a hydrogen atom, and diesters in which both R1 and R2 in the above structural formula are bonded with an acyl group derived from a fatty acid.
• R1 and R2 are the same or different and represent a hydrogen atom or an acyl group derived from a fatty acid, excluding the case where both R1 and R2 are hydrogen atoms (H).
• the acyl group derived from a fatty acid is not particularly limited, and is, for example, a residue obtained by removing a hydroxyl group (OH) from a saturated or unsaturated fatty acid having 8 to 22 carbon atoms.
• the residue obtained by removing a hydroxyl group (OH) from a saturated or unsaturated fatty acid having 8 to 22 carbon atoms can be rephrased as an R 1 CO group or an R 2 CO group.
• R 1 and R 2 may be the same or different, and may be an alkyl group having 7 to 21 carbon atoms, an alkenyl group having 7 to 21 carbon atoms, or an alkynyl group having 7 to 21 carbon atoms.
• saturated fatty acids having 8 to 22 carbon atoms examples include caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, eicosanoic acid (arachidic acid), behenic acid, etc.
• Examples of the unsaturated fatty acids having 8 to 22 carbon atoms include palmitoleic acid, oleic acid, erucic acid, linoleic acid, ⁇ -linolenic acid, and arachidonic acid.
• the acyl group derived from the fatty acid is preferably an acyl group derived from a saturated fatty acid having 12 to 20 carbon atoms, and more preferably a lauroyl group, a myristoyl group, a palmitoyl group, an eicosanoyl group, etc.
• Examples of the fatty acid monoester of capsanthin include capsanthin monomyristate and capsanthin monoeicosanoate.
• Examples of the fatty acid diester of capsanthin include capsanthin dilaurate, capsanthin lauryl myristate, capsanthin dimyristate, capsanthin myristyl palmitate, and capsanthin dipalmitate.
• Capsanthin and/or its fatty acid ester may be any of naturally derived substances such as natural extracts, chemically synthesized substances, and substances derived from microorganisms by microbial fermentation. Capsanthin and/or its fatty acid ester may be commercially available products (Carote Nature, etc.). From the viewpoint of being able to obtain capsanthin and/or its fatty acid ester without relying on chemical means such as chemical synthesis, naturally derived substances are preferred, and plant derived substances are more preferred.
• Capsanthin and/or its fatty acid esters are contained in the red fruits of Capsicum plants of the Solanaceae family (red chili peppers, red paprika, red bell peppers, etc.).
• capsanthin and/or its fatty acid esters are known to be hydrophobic natural pigments contained in the red fruits of Capsicum plants of the Solanaceae family (hereinafter, sometimes referred to as "pigments of Capsicum plants of the Solanaceae family").
• hydrophobicity includes the meaning of lipophilicity or fat-solubility.
• Capsanthin and/or its fatty acid esters which are the active ingredients of the environmental stress resistance improving agent of the present invention, can be obtained from plants containing such capsanthin and/or its fatty acid esters.
• the capsanthin and/or its fatty acid esters are preferably derived from Capsicum plants of the Solanaceae family.
• the pigments of Capsicum plants of the Solanaceae family can be commercially available products (manufactured by San-Ei Gen F.F.I. Co., Ltd., etc.).
• capsanthin and/or its fatty acid ester which is an active ingredient of the environmental stress resistance improving agent of the present invention, from a plant containing capsanthin and/or its fatty acid ester, will be described in detail.
• Plant-derived substances include the plant itself and plant extracts.
• examples of the plant body itself include fruits (red chili peppers, red paprika, red bell peppers, etc.) containing capsanthin and/or its fatty acid esters that are dried and crushed, and further, the dried and crushed materials may be made into a powder, granules, slurry, or paste form.
• the plant extract can be obtained by immersing or contacting a fruit containing capsanthin and/or its fatty acid ester (a plant containing a red pigment such as red pepper, red paprika, or red bell pepper) in an extraction solvent to perform an extraction treatment.
• Capsanthin and/or its fatty acid ester is preferably an extract extracted from at least one plant selected from the group consisting of red paprika, red pepper, and red bell pepper.
• the extraction treatment can be carried out either under normal pressure or under pressure.
• the extraction temperature is usually in the range of 0°C to 150°C, preferably in the range of room temperature to 120°C.
• the extraction time is usually 1 minute to 24 hours, preferably 1 minute to 3 hours.
• the solvent used for the extraction is not particularly limited, and examples thereof include water; acids or bases such as an aqueous hydrochloric acid solution, an aqueous sulfuric acid solution, an aqueous acetic acid solution, and an aqueous sodium hydroxide solution; alcohols such as methanol, ethanol, isopropanol, polyethylene glycol, glycerin, and sorbitol; ketones such as acetone and dimethyl ketone; ethyl acetate, toluene, diethyl ether, methylene chloride, dimethyl sulfoxide, chloroform, hexane, and mixtures thereof.
• acids or bases such as an aqueous hydrochloric acid solution, an aqueous sulfuric acid solution, an aqueous acetic acid solution, and an aqueous sodium hydroxide solution
• alcohols such as methanol, ethanol, isopropanol, polyethylene glycol, glycerin, and sorbi
• the extraction treatment is not particularly limited, but specifically, solvent extraction treatment, subcritical water extraction treatment, heat extraction treatment, pressurized extraction treatment, pressurized hot water extraction treatment, extraction treatment using a Soxhlet extractor, supercritical extraction treatment, ultrasonic extraction treatment, extraction treatment by enzymatic decomposition, etc. can be applied.
• the solvent extraction process refers to an extraction process in which plant material is immersed in a suitable solvent as described above and left to stand for a certain period of time.
• the processing temperature can be room temperature or heated to a temperature below the boiling point of the solvent. It is also possible to promote extraction by stirring the solvent and/or pressurizing the plant material during immersion.
• the solvent extract according to the embodiment of the present invention is obtained by immersing the object to be extracted, such as fruits such as red chili peppers, red paprika, and red bell peppers, in dilute hydrochloric acid heated to 50°C to 70°C for 10 minutes to 24 hours, preferably 30 minutes to 6 hours, and more preferably 1 hour to 3 hours.
• the hot extraction process refers to an extraction process that uses a water-containing solvent at 50°C or higher.
• the hot extract according to the embodiment of the present invention is obtained by contacting or immersing the object to be extracted, such as fruits such as red chili peppers, red paprika, and red bell peppers, in a water-containing solvent at 50°C or higher, and heating or boiling for 3 minutes to 24 hours, preferably 3 minutes to 3 hours, and more preferably 5 minutes to 60 minutes.
• Subcritical water extraction refers to an extraction process that uses a solvent in a subcritical state under conditions of temperature and pressure lower than the critical point (the critical point of water is 22 MPa, 374°C).
• the subcritical water extract according to the embodiment of the present invention is obtained by placing a water solvent or a solvent mixture of water and a hydrophilic solvent such as methanol or acetic acid in a subcritical state, and extracting the subject of extraction, such as fruits such as red chili peppers, red paprika, and red bell peppers, for 1 minute to 24 hours, preferably 1 minute to 60 minutes, and more preferably 3 minutes to 10 minutes.
• the subcritical conditions are preferably a pressure of 0.2 MPa to 15 MPa and a temperature of 100°C to 200°C.
• the plant extract obtained as described above can be concentrated using a rotary evaporator or the like to obtain a plant concentrate containing capsanthin and/or its fatty acid esters. Concentration may be performed using a centrifugal evaporator or freeze-drying.
• the concentration ratio is preferably 5 to 30 times, and more preferably 7.5 to 20 times. A concentration ratio of 5 times or more reduces the amount of environmental stress resistance enhancer added to the plant, improving handling and tending to reduce transportation costs, while a concentration ratio of 30 times or less tends to reduce the amount of environmental stress resistance enhancer precipitated and to be easier to handle.
• the plant extract obtained as described above is obtained in a liquid state extracted in an extraction solvent, but a base or acid may be added to this solvent to perform neutralization treatment.
• the plant extract may be further processed using a vacuum concentration method or a gel adsorption method to obtain a liquid with an increased concentration of capsanthin and/or its fatty acid esters.
• the plant extract may also be processed into a powder by using a spray dryer, a vacuum dryer, or the like.
• the powdered extract may also be dissolved in a solvent other than the extraction solvent to facilitate handling of the plant extract or to increase storage stability.
• capsanthin and/or its fatty acid esters contained in the plant extract it is possible to obtain a plant extract containing high-purity capsanthin and/or its fatty acid esters by adsorption purification using ion exchange resins, silica gel, activated carbon, etc., or purification by column chromatography, recrystallization, etc.
• capsanthin and/or its fatty acid esters are hydrophobic components, so when the plant extract is fractionated by column chromatography, they are included in the hydrophobic fraction (fat-soluble fraction, lipophilic fraction).
• the form of the pigment from the Capsicum plant of the Solanaceae family is not particularly limited, and may be, for example, a liquid such as an emulsion or suspension, a slurry, a paste, or a powder or granules obtained by pulverizing a solid pigment in a conventional manner.
• the pigment of the Capsicum plant of the Solanaceae family may be an extract extracted from a Capsicum plant of the Solanaceae family by a conventional method, as described above (hereinafter, may be referred to as "Capsicum plant extract of the Solanaceae family").
• the pigment of the Capsicum plant of the Solanaceae family may be a commercially available product (manufactured by San-Ei Gen F.F.I. Co., Ltd., etc.).
• the solanaceae Capsicum plant extract referred to here includes, for example, one obtained by crushing a solanaceae Capsicum plant, mixing it with an organic solvent and/or water, and extracting it.
• the environmental stress resistance improver of the present invention may be a plant extract or a pigment of a Capsicum plant of the Solanaceae family obtained as described above, but may be a plant extract that has been subjected to a secondary treatment, as long as the activity of the active ingredient capsanthin and/or its fatty acid ester is not lost and there is no adverse effect on the plant to which it is applied.
• the environmental stress resistance improver of the present invention may be not only a plant extract, but also capsanthin and/or its fatty acid ester obtained by chemical synthesis, or capsanthin and/or its fatty acid ester obtained by microbial fermentation.
• the environmental stress resistance improver of the present invention may be processed into a powder, granules, suspension, etc.
• paprika pigment or capsicum pigment which are registered as food additives in various countries around the world and are available at low cost.
• Commercially available products manufactured by San-Ei Gen F.F.I. Co., Ltd., etc. can be used as the paprika pigment and capsicum pigment.
• the content ratio of the above-mentioned active ingredient in the environmental stress resistance improver can be appropriately determined depending on the dosage form, application mode, etc. described below.
• the concentration of capsanthin and/or its fatty acid ester in the environmental stress resistance improver of the present invention is usually 0.001 to 1,000,000 mg/L, preferably about 0.3 to 30,000 mg/L, and more preferably about 1 to 3,000 mg/L. Even when applied at such a low concentration, the environmental stress resistance improver of the present invention can improve the environmental stress resistance of the plant to be protected.
• the application of the environmental stress resistance improver of the present invention to the plant body can be performed only once throughout the entire cultivation period of the plant body to sufficiently improve the environmental stress resistance of the plant to be protected, but it is also preferable to apply it multiple times in order to sufficiently increase the environmental stress resistance.
• the content of capsanthin and/or its fatty acid ester in the paprika colorant used in the present invention is not particularly limited, and is, for example, usually 0.001 to 3,000,000 mg, preferably about 0.1 to 30,000 mg, and more preferably about 1 to 3,000 mg per 100 g of paprika colorant.
• the formulation of the environmental stress resistance improving agent of the present invention is not particularly limited as long as it is an agriculturally acceptable formulation (use form), and examples include flowable formulations such as wettable powders, wettable granules, dry flowable agents, water-soluble agents, emulsions, liquid agents (also called liquid-like or liquid), oil agents, paste agents, dispersants, water suspensions, water emulsions, etc., capsules, powders, granules, fine granules, granules, baits, tablets, sprays, aerosols, etc.
• various additives (auxiliaries) conventionally used in the technical field of agricultural and horticultural agents can be used as appropriate.
• the environmental stress resistance improving agent of the present invention may consist only of the above-mentioned active ingredients, but in addition to the above-mentioned active ingredients, it may also contain various fertilizer components, additives, etc. depending on the formulation, usage form, application mode, etc. described below.
• the environmental stress resistance improver composition of the present invention further contains a fertilizer component and/or an additive in addition to the above-mentioned environmental stress resistance improver.
• a fertilizer component can be further used together with capsanthin and/or its fatty acid ester.
• the present invention encompasses an environmental stress resistance improver composition that further contains a fertilizer component in addition to the environmental stress resistance improver.
• the environmental stress resistance improver composition can be rephrased as a composition for improving environmental stress resistance, a composition for improving environmental stress resistance, etc.
• the fertilizer components are not particularly limited as long as they are agriculturally acceptable fertilizer components, and examples thereof include N (nitrogen source), P (phosphate source), K (potash source), Ca (lime source), Mg (magnesium source), S (sulfur source), B (boron source), Fe (iron source), Mn (manganese source), Cu (copper source), Zn (zinc source), Mo (molybdenum source), and Si (silicic acid source), as well as inorganic substances (inorganic fertilizers) and organic substances (organic fertilizers) that are the supply sources of these.
• Examples of such inorganic substances include ammonium nitrate, potassium nitrate, ammonium sulfate, ammonium chloride, ammonium phosphate, urea, ammonium carbonate, potassium phosphate, lime superphosphate, potassium sulfate, potassium chloride, lime nitrate, slaked lime, lime carbonate, magnesium sulfate, magnesium hydroxide, magnesium carbonate, boric acid, manganese sulfate, manganese chloride, chelated iron, ferric chloride, chelated copper, chelated zinc, and ammonium molybdate.
• Examples of organic matter include chicken manure, cow manure, bark compost, seaweed, plant extracts, vitamins (vitamin B1, vitamin C, etc.), amino acids (glycine, glutamic acid, sodium glutamate, etc.) and their salts, etc.
• the concentrations of the fertilizer components in the environmental stress resistance improving composition are, when applied to a plant body, preferably 0 to 5000 mg/L for the N component, P component, and K component, more preferably 0 to 1000 mg/L, and even more preferably 0 to 500 mg/L for each of the N component, P component, and K component, when sprayed on the leaves.
• concentrations of the fertilizer components in the environmental stress resistance improving composition are, when applied to a plant body, preferably 0 to 5000 mg/L for the N component, P component, and K component, more preferably 0 to 1000 mg/L, and even more preferably 0 to 500 mg/L for each of the N component, P component, and K component.
• the concentration of all the fertilizer components combined is preferably 0.01 to 10000 mg/L, more preferably 10 to 5000 mg/L, and even more preferably 50 to 2000 mg/L.
• the concentration of all fertilizer components combined is preferably 0.01 to 10,000 mg/L, more preferably 10 to 5,000 mg/L, and even more preferably 50 to 2,000 mg/L, when applied underground in soil and hydroponic cultivation.
• the environmental stress resistance improver composition of the present invention may further contain an additive.
• the present invention includes an environmental stress resistance improver composition that contains, in addition to the environmental stress resistance improver, further additives.
• the present invention also includes an environmental stress resistance improver composition that contains, in addition to the environmental stress resistance improver, a fertilizer component and an additive.
• the additives are not particularly limited as long as they are agriculturally acceptable additives, and examples thereof include antioxidants, carriers, surfactants, thickeners, extenders, binders, preservatives, pH adjusters, evaporation inhibitors, and precipitation inhibitors.
• the concentration thereof can be appropriately adjusted within a range that does not impair the effectiveness of capsanthin or its fatty acid ester.
• the environmental stress resistance improving composition of the present invention may further contain other known active compounds, such as insecticides, acaricides, nematicides, fungicides, synergists, and plant regulators.
• a surfactant As the additive, it is possible to improve the wettability, adhesion, permeability, rain resistance, etc. of capsanthin and/or its fatty acid ester on the plant surface, etc.
• nonionic surfactants those that dissolve in water, such as nonionic surfactants, cationic surfactants, amphoteric surfactants and anionic surfactants
• nonionic surfactants include sorbitan fatty acid esters, polyoxyalkylene sorbitan fatty acid esters, polyoxyalkylene fatty acid esters, glycerin fatty acid esters, polyoxyalkylene glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyalkylene polyglycerin fatty acid esters, sucrose fatty acid esters, resin acid esters, polyoxyalkylene resin acid esters, polyoxyalkylene alkyl ethers, polyoxyalkylene alkylphenyl ethers, alkyl (poly)glycosides, polyoxyalkylene alkyl (poly)glycosides, etc.
• nonionic surfactants preferred are ether group-containing nonionic surfactants that do not contain nitrogen atoms, and ester group-containing nonionic surfactants.
• ester group-containing nonionic surfactants that contain oxyalkylene groups, such as polyoxyalkylene sorbitan fatty acid esters, polyoxyalkylene fatty acid esters, polyoxyalkylene glycerin fatty acid esters, and polyoxyalkylene polyglycerin fatty acid esters; and ether group-containing nonionic surfactants that do not contain nitrogen atoms and have a sugar skeleton, such as alkyl (poly)glycosides.
• anionic surfactants include carboxylic acid surfactants, sulfonic acid surfactants, sulfate surfactants, and phosphate surfactants.
• carboxylic acid surfactants include fatty acids having 6 to 30 carbon atoms or their salts, polyvalent carboxylates, polyoxyalkylene alkyl ether carboxylates, polyoxyalkylene alkyl amide ether carboxylates, rosin acid salts, dimer acid salts, polymer acid salts, tall oil fatty acid salts, etc.
• sulfonic acid surfactants include alkyl benzene sulfonates, alkyl sulfonates, alkyl naphthalene sulfonates, naphthalene sulfonates, diphenyl ether sulfonates, condensate salts of alkyl naphthalene sulfonic acid, and condensate salts of naphthalene sulfonic acid.
• sulfate surfactants include alkyl sulfate salts, polyoxyalkylene alkyl sulfate salts, polyoxyalkylene alkyl phenyl ether sulfate salts, tristyrenated phenol sulfate salts, polyoxyalkylene distyrenated phenol sulfate salts, and alkyl polyglycoside sulfate salts.
• phosphate surfactants include alkyl phosphate salts, alkylphenyl phosphate salts, polyoxyalkylene alkyl phosphate salts, polyoxyalkylene alkylphenyl phosphate salts, etc.
• salts examples include metal salts (Na, K, Ca, Mg, Zn, etc.), ammonium salts, alkanolamine salts, aliphatic amine salts, etc.
• anionic surfactants include carboxylic acid surfactants and phosphate surfactants.
• amphoteric surfactants include amino acid surfactants, betaine surfactants, imidazoline surfactants, and amine oxide surfactants.
• amino acid surfactants include acylamino acid salts, acyl sarcosinate salts, acyloylmethylaminopropionate salts, alkylaminopropionate salts, and acylamidoethyl hydroxyethyl methyl carboxylate salts.
• betaine surfactants include alkyl dimethyl betaines, alkyl hydroxyethyl betaines, acylamidopropyl hydroxypropyl ammonia sulfobetaines, acylamidopropyl hydroxypropyl ammonia sulfobetaines, and ricinoleic acid amidopropyl dimethyl carboxymethyl ammonia betaines.
• imidazoline surfactants include alkyl carboxymethyl hydroxyethyl imidazolinium betaines and alkyl ethoxy carboxymethyl imidazolium betaines.
• amine oxide surfactants include alkyl dimethyl amine oxides, alkyl diethanol amine oxides, and alkyl amidopropyl amine oxides. The above surfactants may be used alone or in combination of two or more.
• the concentration of the surfactant in the environmental stress resistance improver composition is preferably 0.01 to 1000 mg/L, more preferably 0.03 to 300 mg/L, and even more preferably 0.1 to 100 mg/L, when applied to the plant body in foliar spraying. Also, the concentration of the surfactant in the environmental stress resistance improver composition is preferably 0.01 to 1000 mg/L, more preferably 0.03 to 300 mg/L, and even more preferably 0.1 to 100 mg/L, when applied to the above-ground parts of the plant body in soil and hydroponic cultivation.
• the mass ratio of capsanthin and/or its fatty acid ester to surfactant is preferably capsanthin and/or its fatty acid ester:surfactant of 1:0.001-1000, more preferably 1:0.01-100, and even more preferably 1:0.1-10.
• an environmental stress resistance enhancer containing capsanthin and/or its fatty acid ester to the above-ground or underground parts of a plant, plant growth is promoted under normal conditions, and stress resistance is improved to promote plant growth even in stressful environments.
• Stresses to which the environmental stress tolerance improver of the present invention improves tolerance include high temperature, dryness, low temperature, heavy rain, ultraviolet light, salt, etc.
• Examples of environmental stress tolerance improved by the environmental stress tolerance improver include high temperature stress tolerance, dryness stress tolerance, and salt stress tolerance.
• high temperature stress tolerance refers to tolerance to high temperatures that exceed the optimum growth temperature of the target plant.
• Dryness stress tolerance refers to tolerance to a state without water for a certain period of time.
• Salt stress tolerance refers to tolerance to high salt concentrations in the plant's growth environment, including moisture in contact with the soil or stems and leaves.
• improved environmental stress resistance means that the growth status of the plant (fresh weight of the plant, degree of elongation, etc.), the condition as evaluated visually from the appearance, etc. are judged to be in a better condition than the plants in the control group.
• the target plant of the environmental stress resistance improver of the present invention is not particularly limited.
• Useful plants that can use the environmental stress resistance improver of the present invention are not particularly limited, and include, for example, tomato, eggplant, green pepper, chili pepper, potato, etc. of the Solanaceae family; rice, barley, wheat, rye, corn, sugarcane, etc. of the Poaceae family; soybean, adzuki bean, broad bean, pea, kidney bean, peanut, etc. of the Leguminosae family; apple, pear, peach, plum, cherry, strawberry, rose, etc. of the Rosaceae family; cabbage, broccoli, Chinese cabbage, radish, turnip, komatsuna, etc.
• Lamiaceae Amaranthaceae such as spinach and sugar beet; Apiaceae such as carrot and parsley; Asteraceae such as lettuce, burdock, and chrysanthemum; Convolvulaceae such as sweet potato; other fruits such as citrus fruits, grapes, chestnuts, almonds, and bananas; root vegetables such as taro and lotus root; processing crops such as cotton, hemp, hops, olives, rubber, coffee, and tea; pasture grasses such as orchard grass, sorghum, timothy, clover, and alfalfa; turf grasses such as Korean grass and bentgrass; ornamental crops such as pepper and ginger; floral plants such as carnations and orchids; garden trees such as ginkgo, cherry trees, and Japanese laurel; forest trees such as Abies sachalinensis, Ezo spruce, pines, Japanese cypress, cedar, and cypress. Furthermore, the target for improving environmental stress resistance is not limited
• the application mode of the environmental stress resistance enhancer or environmental stress resistance enhancer composition of the present invention is not particularly limited as long as it is a known mode of use for pesticides or fertilizers (or a mode to be developed in the future), and examples of such modes include scattering, spraying, dripping, coating, adding, immersion, mixing or dissolving in the plant growth environment (in soil, water, solid medium, liquid medium, culture solution, etc.).
• the present invention includes a method for improving environmental stress tolerance of a plant, which comprises applying the above-mentioned environmental stress tolerance improver or environmental stress tolerance improver composition to a plant or to the soil or culture solution in which the plant grows.
• a method for improving environmental stress tolerance of a plant which comprises applying the above-mentioned environmental stress tolerance improver or environmental stress tolerance improver composition to a plant or to the soil or culture solution in which the plant grows.
• the environmental stress resistance improver or environmental stress resistance improver composition of the present invention can be applied to a plant or a part thereof (e.g., a seed, a seedling, or a mature plant body) at any growth stage, including before or after germination. In addition, it can be applied not only to the plant itself, but also to the soil, medium, or culture solution in which the plant grows.
• the environmental stress resistance improver or environmental stress resistance improver composition By applying the environmental stress resistance improver or environmental stress resistance improver composition to a plant at any growth stage or to the soil, medium, or culture solution in which the plant grows, the growth of the plant can be promoted.
• the environmental stress resistance enhancer or the environmental stress resistance enhancer composition may be applied to a plant once or multiple times. Furthermore, when the application period is multiple, the number of applications at each application period may be once or multiple times.
• the present invention includes an agent for improving environmental stress tolerance that activates a gene in a stress response system.
• the gene in the stress response system is not particularly limited, and examples thereof include genes generally known as response indicators to temperature stress, chemical stress, light stress, drought stress, pH stress, salt stress, hypoxic stress, herbicide stress, physical stress, disease stress, etc.
• the environmental stress resistance improving agent of the present invention that activates genes in the stress response system can increase or promote the expression level of genes that contribute to heat resistance (e.g., HSF genes, HSP genes, etc.) by using paprika pigments.
• the HSF genes are an abbreviation for heat shock transcription factors (HSFs), and refer to a group of transcription factors involved in the control of the expression of a group of heat shock proteins.
• HSF gene include HSFA1 (HSFA1a, HSFA1b, HSFA 1d, HSFA1e, etc.), HSFA3, etc.
• HSF gene contributes to the acquisition of high temperature stress resistance, so it is expected that the use of paprika pigment will improve the high temperature stress resistance of crops.
• paprika pigment has an effect of changing the gene expression level in many plants other than tomato (Reference (Non-Patent Document 2): Klaus-Dieter Scharf et al., The plant heat stress transcription factor (Hsf) family: Structure, function and evolution. Biochimica et Biophysica Acta. Volume 1819(2), 2012, Pages 104-119.
• Example 1 (Test to confirm the growth promotion effect of red paprika extract in hydroponic rice cultivation)> Example 1 According to the method described in Patent No. 7317275 (Agri Smile Co., Ltd.), an extract was extracted from red pepper pulp and separated into a hydrophilic fraction and a hydrophobic fraction as described below. First, the stems of 1 kg of red peppers were removed, cut into 1 cm cubes, and then crushed in a mixer. Impurities were removed by filtration using filter cloth and filter paper. The obtained filtrate was concentrated about 10 times using a rotary evaporator to obtain a concentrated solution.
• Comparative Examples 1 and 2 As comparative examples, a test similar to that of Example 1 above was performed, except that the liquid medium was replaced with one to which no hydrophobic fraction was added (Comparative Example 1: untreated), or with a liquid medium to which a hydrophilic fraction was added instead of the hydrophobic fraction (Comparative Example 2).
• Reference Example 1 In Reference Example 1, a test similar to that in Example 1 was carried out, except that instead of the hydrophilic fraction or the hydrophobic fraction, a commercially available high-temperature resistance improving material (material A) was added to the liquid medium at the dilution ratio shown in Table 1. In Reference Example 1, the dilution ratio is shown because the content of the active ingredient in the commercially available product is unknown. The following Reference Examples are calculated using the dilution ratio for the same reason.
• Example 2 A liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask. Then, the hydrophobic fraction (Example 2) was added to the liquid medium at a concentration (volume %) shown in Table 2. Next, rice (Koshihikari) was grown in culture soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivation at 35°C for 6 days, the length of the rice roots was measured and the regenerated root length was calculated.
• OAT House Fertilizer registered trademark
• Comparative Examples 3 and 4 As comparative examples, tests were conducted similarly to Example 2 above, except that the liquid medium was replaced with one to which no hydrophobic fraction was added (Comparative Example 3), or with one to which a hydrophilic fraction was added instead of the hydrophobic fraction (Comparative Example 4).
• Reference Example 2 As Reference Example 2, a test similar to that of Example 2 was carried out, except that instead of the hydrophilic fraction or the hydrophobic fraction, a liquid medium was used to which a commercially available high-temperature resistance improving material (material A) was added at the dilution ratio shown in Table 2. In Reference Example 2, the dilution ratio is shown because the content of the active ingredient in the commercially available product is unknown.
• material A high-temperature resistance improving material
• Example 3 (Test to confirm growth promotion effect in hydroponic rice cultivation)> Example 3
• a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask.
• Capsanthin or paprika pigment was added to the liquid medium to the concentration shown in Table 3.
• the capsanthin concentration in the paprika pigment (manufactured by San-Ei Gen F.F.I. Co., Ltd.) was measured by high performance liquid chromatography at the Japan Food Analysis Center General Incorporated Foundation. As a result, about 221 mg of capsanthin was contained in 100 g of the paprika pigment.
• the measurement conditions are as follows.
• Comparative Example 5 As Comparative Example 5, a test similar to that in Example 3 was carried out, except that a liquid medium containing neither capsanthin nor paprika pigment was used.
• Reference Example 3 As Reference Example 3, a test similar to that of Example 3 was conducted, except that a commercially available high-temperature resistance improving material (material A) was added at the dilution ratio shown in Table 3 instead of the liquid medium. In Reference Example 3, the dilution ratio of the commercially available product is shown since the content of the active ingredient is unknown.
• material A high-temperature resistance improving material
• Example 3 results are shown as a ratio (%) when the result of Comparative Example 5 (untreated area) is set to 100.
• Example 4 (Test for improving high temperature stress resistance in rice hydroponic cultivation)> Example 4
• a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask.
• the above-mentioned capsanthin or paprika pigment was added to the liquid medium to the concentration shown in Table 4.
• rice (Koshihikari) was grown in culture soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivation at 35°C for 6 days, the length of the rice roots was measured and the regenerated root length was calculated.
• Comparative Example 6 As Comparative Example 6, a test similar to that of Example 4 was carried out, except that a liquid medium containing neither capsanthin nor paprika pigment was used.
• Example 4 results are shown as a percentage when the results of Comparative Example 6 (untreated area) are taken as 100.
• Example 5 (Test for improving salt stress tolerance in rice hydroponic cultivation)> Example 5 As Example 5, a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask. Sodium chloride was added to the liquid medium to a final concentration of 120 mM, and capsanthin or paprika pigment was added to the liquid medium to the concentrations shown in Table 3. Next, rice (Koshihikari) was grown in soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivation for six days, the length of the rice roots was measured and the regenerated root length was calculated.
• OAT House Fertilizer registered trademark
• capsanthin or paprika pigment was added to the liquid medium to the concentrations shown in Table 3.
• Comparative Example 7 As Comparative Example 7, a test similar to that of Example 5 was carried out, except that a liquid medium containing neither capsanthin nor paprika pigment was used.
• Reference Example 4 As Reference Example 4, a test similar to that of Example 5 was conducted, except that the liquid medium was changed to one to which a commercially available salt tolerance improving material (material B) was added at the dilution ratio shown in Table 5, instead of capsanthin and paprika pigment.
• material B a commercially available salt tolerance improving material
• Example 5 results of Example 5 and Reference Example 4 are shown as a ratio (%) when the result of Comparative Example 7 (untreated area) is set to 100.
• Example 6 (Test to confirm growth promotion effect in hydroponic rice cultivation)> Example 6 As Example 6, a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask. Paprika pigment was added to the liquid medium to the concentration shown in Table 6. Next, rice (Koshihikari) was grown in soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivation at 25°C for 6 days, the length of the rice roots was measured and the regenerated root length was calculated.
• OAT House Fertilizer registered trademark
• Comparative Example 8 As Comparative Example 8, a test similar to that of Example 6 was carried out, except that a liquid medium containing no paprika pigment was used instead.
• Reference Example 5 In Reference Example 5, a test similar to that of Example 6 was conducted on the paprika pigment, except that the paprika pigment was replaced with a liquid medium containing a commercially available high-temperature resistance improving material (material A) at the dilution ratio shown in Table 1.
• the dilution ratio is shown because the content of the active ingredient in the commercially available product is unknown.
• the following Reference Examples are calculated using the dilution ratio for the same reason. These results are shown in Table 6.
• the results of Example 6 and Reference Example 5 are shown as percentages when the result of Comparative Example 8 (untreated area) is taken as 100.
• Example 7 (Test for improving high temperature stress resistance in rice hydroponic cultivation)> Example 7 As Example 7, a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask. Paprika pigment was added to the liquid medium to the concentration shown in Table 7. Next, rice (Koshihikari) was grown in soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivating the rice at 35°C for 6 days, the length of the rice roots was measured and the regenerated root length was calculated.
• OAT House Fertilizer registered trademark
• Paprika pigment was added to the liquid medium to the concentration shown in Table 7.
• Comparative Example 9 As Comparative Example 9, a test similar to that of Example 7 was carried out, except that a liquid medium containing no paprika pigment was used instead.
• Reference Example 6 As Reference Example 6, a test similar to that of Example 7 above was conducted, except that the paprika pigment was replaced with a liquid medium to which a commercially available high-temperature resistance improving material (material A) was added at the dilution ratio shown in Table 2. These results are shown in Table 7. The results of Example 7 and Reference Example 6 are shown as percentages when the result of Comparative Example 9 (untreated area) is taken as 100.
• Example 8 (Test for improving salt stress tolerance in rice hydroponic cultivation)> Example 8 As Example 8, a liquid medium prepared by diluting OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd. was placed in an Erlenmeyer flask. Sodium chloride was added to the liquid medium to a final concentration of 120 mM, and paprika pigment was added to the liquid medium to a concentration shown in Table 8. Next, rice (Koshihikari) was grown in culture soil, and the roots were cut to a certain length and inserted into an Erlenmeyer flask in groups of five. After cultivation for six days, the length of the rice roots was measured, and the regenerated root length was calculated.
• OAT House Fertilizer registered trademark
• Comparative Example 10 As Comparative Example 10, a test similar to that of Example 8 was carried out, except that a liquid medium containing no paprika pigment was used instead.
• Reference Example 7 As Reference Example 7, a test similar to that of Example 8 above was conducted, except that the paprika pigment was replaced with a liquid medium to which a commercially available salt tolerance improving material (material B) was added at the dilution ratio shown in Table 8. These results are shown in Table 8. The results of Example 8 and Reference Example 7 are shown as percentages when the result of Comparative Example 10 (untreated area) is taken as 100.
• Example 9 (Test for improving drought stress resistance in tomatoes)> Example 9
• tomatoes variety: Ponterosa
• aqueous paprika dye solution prepared to the concentration shown in Table 9. After removing excess water, water supply was completely withheld and water was withheld from the untreated area, and water was resupplied when the untreated area withered and died, allowing the plants to recover. The survival rate was calculated from the number of surviving tomato plants.
• Comparative Example 11 As Comparative Example 11, a test similar to that of Example 9 was carried out except that water to which no paprika pigment or the like was added was used.
• Reference Example 8 As Reference Example 8, a test similar to that of the paprika pigment of Example 9 was carried out, except that an aqueous solution containing a commercially available indigo pigment added to the concentration shown in Table 9 was used instead of the paprika pigment. These results are shown in Table 9 and Figure 1.
• Figure 1 is a photograph ( Figures 1a and 1b) showing the state of the tomato seedlings after the test.
• the left side of Figure 1a is a photograph of Comparative Example 11 (untreated), and the right side of Figure 1a is a photograph of Example 9 (paprika pigment).
• the left side of Figure 1b is a photograph of Comparative Example 11 (untreated), and the right side of Figure 1b is a photograph of Reference Example 8 (indigo pigment).
• Example 10 (Test for improving drought stress tolerance in wheat)> Example 10
• wheat variety: Norin 61
• a hydroponic cultivation system registered trademark
• Example 10 In Example 10, wheat (variety: Norin 61) was sown in a cultivation tank in a greenhouse, irrigated with tap water using a hydroponic cultivation system (registered trademark), and cultivated until the heading stage. Next, after wheat heading, irrigation was continued in the fully irrigated area, and water was stopped for 7 days in the drought stress area. Then, after water supply was stopped and water supply was resumed, an aqueous paprika pigment solution prepared to the concentration shown in Table 10 was sprayed on the foliage of the fully irrigated area and the drought stress area. Then, the number of ears per plant was investigated 132 days after sowing in the fully irrigated area and 137 days after sowing in the drought stress area.
• Comparative Example 12 As Comparative Example 12, a test similar to that of Example 10 was carried out, except that water (containing 100 ppm of Tween (registered trademark; hereinafter omitted) 20) to which no paprika colorant or the like was added was used.
• Reference Example 9 As Reference Example 9, a test similar to that of the paprika pigment in Example 10 was conducted, except that an aqueous solution containing a commercially available indigo pigment added to the concentration shown in Table 10 was used instead of the paprika pigment. These results are shown in Table 10. The results of Example 10 and Reference Example 9 are shown as percentages when the result of Comparative Example 12 (untreated area) is taken as 100.
• the paprika pigment of Example 10 increased the number of ears compared to the water treatment area under both sufficient irrigation conditions and drought stress conditions, and also showed growth promotion effects and drought stress resistance improvement effects.
• Example 11 (Test for improving drought stress tolerance in wheat)> Example 11
• wheat variety: Chikugoizumi
• Example 11 wheat (variety: Chikugoizumi) was sown in 1/5000a Wagner pots and cultivated at a density of 2 plants per pot.
• all pots were irrigated with the same amount of tap water until the wheat head emerged, and irrigation with a chemical solution was started after the wheat head emerged. In this test, irrigation was limited to about half the usual amount to induce drought stress.
• the aboveground parts of the wheat were harvested and the grain weight per plant was investigated. A 30 ppm aqueous solution of paprika coloring was used as the chemical solution.
• Comparative Example 13 As Comparative Example 13, a test similar to that of the paprika colorant in Example 11 was carried out, except that water was used for irrigation after ear emergence.
• Reference Examples 10 and 11 As Reference Example 10, a test similar to that of the paprika colorant of Example 11 was conducted except that a commercially available salt tolerance improving material (material B) was used at the dilution ratio shown in Table 11. Also, as Reference Example 11, a test similar to that of the paprika colorant of Example 11 was conducted except that potassium acetate was used at the concentration shown in Table 11. These results are shown in Table 11. The results of Example 11, Reference Example 10 and Reference Example 11 are shown as percentages when the result of Comparative Example 13 (untreated area) is taken as 100.
• a commercially available salt tolerance improving material material B
• potassium acetate was used at the concentration shown in Table 11.
• the paprika color of Example 11 was shown to increase wheat grain yield and improve drought stress resistance under conditions where the wheat was cultivated with a significantly smaller amount of irrigation than usual. Therefore, in the present invention, the paprika color containing capsanthin had the effect of increasing grain yield under drought stress.
• Example 12 (Pod number increase test for kidney beans)> Example 12
• kidney beans variety: Hatsumidori 2
• Six kidney bean plants were used in each test plot. After flowering, the plants were sprayed twice in total with a spray bottle at 5 mL per plant with a paprika pigment solution prepared to the concentration shown in Table 7. The plants were harvested when the primary leaves began to yellow and fall, and the number of pods was counted for pods with a pod length of 6 cm or more.
• Comparative Example 14 As Comparative Example 14, a test was carried out similarly to that of Example 12, except that water was used instead of the aqueous paprika pigment solution.
• Reference Example 12 As Reference Example 12 (positive control), a test similar to that of Example 12 above was conducted, except that a commercially available seaweed extract material (material C) was used at the dilution ratio shown in Table 12. The results of these evaluations are shown in Table 12. The results of Example 12 and Reference Example 13 are shown as percentages when the result of Comparative Example 14 (untreated area) is taken as 100.
• the paprika color of Example 12 increased the number of pods of kidney beans at 0.003 ppm and 30 ppm. Therefore, in the present invention, the paprika color containing capsanthin had the effect of increasing the number of pods under normal conditions.
• Example 13 (Test for improving high temperature stress resistance in tomatoes)> Example 13 As Example 13, on June 3, 2022, two tomatoes (variety: Karen) were planted in a planter and grown in a vinyl greenhouse up to the 14th stage. One week after planting, a paprika pigment aqueous solution adjusted to the concentration shown in Table 8 was periodically sprayed on the entire tomato plant at 100 mL/plant from the time of the first stage flowering, and the number of flowers and fruits at each stage were measured. During the cultivation period, the plant was appropriately irrigated and fertilized with Tank Mix (registered trademark) A & B formulation manufactured by OAT Agrio Co., Ltd.
• Tank Mix registered trademark
• Comparative Example 15 As Comparative Example 15, the same test as that of the above paprika pigment was carried out, except that water was used instead of the aqueous paprika pigment solution. The results of the number of flowers and the number of fruits in the tier where the average temperature in the greenhouse was 30° C. or higher are shown in Table 13. Note that the average temperature in the greenhouse refers to the average temperature including the night temperature, and an environment where the average temperature is 30° C. or higher can be said to be a high temperature condition for tomato cultivation. The results of Example 13 are shown as a ratio (%) when the result of Comparative Example 15 (untreated area) is set to 100.
• the paprika colorant of Example 13 increased the number of tomato flowers and fruit set at the stage where flowering and fruit set occurred under high temperature conditions (when the average temperature in the greenhouse was 30°C or higher) compared to the untreated group of Comparative Example 15. Therefore, in the present invention, the paprika colorant containing capsanthin had an effect of increasing or promoting the number of flowers and the number of fruit set under high temperature stress.
• Comparative Example 16 As Comparative Example 16 (untreated group), the same test as in Example 14 was carried out, except that water was used instead of the aqueous paprika pigment solution. The evaluation results are shown in Table 14.
• the paprika colorant of Example 14 promoted flower bud differentiation of strawberry seedlings grown outdoors in the summer under high temperature conditions, compared to the untreated group of Comparative Example 16. Therefore, in the present invention, the paprika colorant containing capsanthin had the effect of promoting flower bud differentiation under high temperature stress.
• Example 15 (Test for improving resistance to high temperature stress in strawberries)> Examples 15 to 17 From July 19, 2023, strawberry seedlings grown outdoors were sprayed with a paprika dye solution adjusted to a specified concentration on the entire plant at one-week intervals from September 1 until transplantation, and at two-week intervals after transplantation into pots. Six plants were set up in each area, and the number of days from transplantation to the first flowering was counted. During the cultivation period, the plants were appropriately irrigated and fertilized with a diluted solution of OAT House Fertilizer (registered trademark) A formulation manufactured by OAT Agrio Co., Ltd.
• OAT House Fertilizer registered trademark
• Comparative Examples 17 to 19 As Comparative Examples 17 to 19 (untreated groups), the same tests as those for the above-mentioned paprika pigment were carried out, except that water was used instead of the aqueous paprika pigment solution. The evaluation results are shown in Table 15.
• Example 16 (Test for improving high temperature stress resistance in komatsuna)>
• Example 18 A 10 ppm solution of paprika pigment was sprayed on the leaves of komatsuna at the 4.5 leaf stage, and the plants were grown under the following temperature conditions: 17 hours of light (7:00-24:00), 7 hours of darkness (0:00-7:00), and temperatures of 25°C from 0:00 to 5:00, 28°C from 5:00 to 6:00, 31°C from 6:00 to 7:00, 35°C from 7:00 to 16:00, 32°C from 16:00 to 17:00, 29°C from 17:00 to 18:00, and 25°C from 18:00 to 24:00.
• the komatsuna was sprayed a second time, and the plants were grown for one week, after which a harvest survey was conducted. The temperature conditions during growth were set with reference to the temperature inside the greenhouse in summer.
• Comparative Example 20 As Comparative Example 20 (untreated group), the same test as that of the above paprika pigment was carried out, except that water was used instead of the aqueous paprika pigment solution. The evaluation results are shown in Table 16.
• the treatment area of the paprika colorant of Example 18 increased the fresh weight of the aboveground part, which is the edible part of Komatsuna grown under high temperature conditions, by about 9% compared to the untreated area of Comparative Example 20. Therefore, in the present invention, the paprika colorant containing capsanthin had the effect of increasing the fresh weight of the aboveground part under high temperature stress.
• Examples 19 to 21 Using the measurement method described below, expression levels of high temperature stress marker genes SIHSFA1a and SIHSFA3 were analyzed using a paprika pigment-sprayed area grown under no high temperature stress (unheated) conditions, an untreated area (water-sprayed area) grown under high temperature stress (heated) conditions, and a paprika pigment-sprayed area grown under high temperature stress (heated) conditions as examples.
• the expression level in the water-sprayed group grown without high temperature stress (Comparative Example 21) was set at 1.0 and used as the standard.
• Example 21 No high temperature stress (non-heated) and non-treated area (water sprayed area) (Comparative Example 21), - No high temperature stress (non-heated) and paprika dye sprayed area (Example 19), - With high temperature stress (heating) and no treatment (water spraying) (Example 20), - High temperature stress (heating) and paprika dye spraying area (Example 21)
• RNA extraction and cDNA synthesis The extraction of total RNA from the leaf tissue of the plant body and the removal of genomic DNA were performed using RNAiso Plus (manufactured by Takara Bio Inc.). 30 ⁇ L of RNase-free Water (manufactured by Takara Bio Inc.) was used to elute the RNA. The concentration of the RNA solution was determined using a microspectrophotometer (NANODROP (registered trademark), manufactured by Thermo Fisher Scientific Inc.), and it was confirmed that there was no problem with the quality of the RNA sample. Then, a reverse transcription reaction was performed using ReverTra Ace (registered trademark) (manufactured by Toyobo Co., Ltd.), and cDNA was synthesized.
• NANODROP registered trademark
• ReverTra Ace registered trademark
• RT-qPCR The enzyme for RT-qPCR was KAPA SYBR Fast qPCR kit (Universal qPCR kit) (manufactured by Nippon Genetics Co., Ltd.), and 1 ⁇ L of cDNA solution was added to 20 ⁇ L of reaction solution. PCR was performed using a qRT-PCR system (Thermal Cycler Dice Real Time System III). After data acquisition, the expression levels of each gene were compared by the Comparative CT method ( ⁇ CT method). The expression level of the ACT gene was used as a reference and standardized with the value of the untreated area (water sprayed area) that was not subjected to high temperature stress.
• ⁇ CT method Comparative CT method
• Example 22 On June 1, 2023, 100 kg/10a of superphosphate (manufactured by Asahi Agria Co., Ltd.) and chemical fertilizer (manufactured by JA (Agricultural Cooperative Association), 16% nitrogen, 16% phosphoric acid, 16% potassium) were applied as nitrogen (N) to give 15 kg/10a, and spinach seeds (variety: Justice) were sown at 5 cm between plants, 15 cm between rows, and a sowing depth of 1 cm. 20 plants per row, 80 plants per test area. During the growth period, the soil surface was checked and watered appropriately. The above Formulation Example 4 was sprayed periodically over the entire plant. A yield survey was conducted on July 11, 2023.
• Comparative Example 22 As a comparative example, a test similar to that for the solution containing paprika pigment was carried out, except that water was used instead of the solution used in Example 22 (Formulation Example 4).
• Reference Example 13 As a reference example (positive control), a test similar to that of Example 22 (Formulation Example 4) was conducted, except that a commercially available high temperature resistance improving material (Material A) was used at the dilution ratio shown in the table. The results are shown in the following Table 18. The results of Example 22 and Reference Example 13 are shown as percentages when the result of Comparative Example 22 (untreated) is taken as 100.

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